Cloning, expression and purification of a polytopic antigen comprising of surface antigens of Toxoplasma gondii
,
Abstract
Background and Objectives: Polytopic antigens are recently applied for replacing crude antigens, for control of infectious agents. The surface of the Toxoplasma is covered with immunogenic antigens namely surface antigens (SAGs). These antigens possess several immunogenic epitopes, inducing immune responses.
Materials and Methods: In this study, a DNA construct comprising of sequences encoding epitopes from SAG1, 2 and 3 was designed and cloned into pET28a expression vector and subsequently expressed and purified, using Ni-NTA column.
Results: The SDS-PAGE and Western blotting analysis showed that polytopic genes were successfully expressed and purified.
Conclusion: The surface antigenic protein of T. gondii can be applied in the future epitope-based applications.
Hide G. Role of vertical transmission of Toxoplasma gondii in prevalence of infection. Expert Rev Anti Infect Ther 2016;14:335-344.
Smith JE. A ubiquitous intracellular parasite: the cellular biology of Toxoplasma gondii. Int J Parasitol 1995;25:1301-1309.
Lekutis C, Ferguson DJ, Grigg ME, Camps M, Boothroyd JC. Surface antigens of Toxoplasma gondii: variations on a theme. Int J Parasitol 2001;31:1285-1292.
Wang Y, Yin H. Research progress on surface antigen 1 (SAG1) of Toxoplasma gondii. Parasit Vectors 2014;7:180.
Macedo AG, Cunha JP, Cardoso TH, Silva MV, Santiago FM, Silva JS, et al. SAG2A protein from Toxoplasma gondii interacts with both innate and adaptive immune compartments of infected hosts. Parasit Vectors 2013;6:163.
Jacquet A, Coulon L, De Neve J, Daminet V, Haumont M, Garcia L, et al. The surface antigen SAG3 mediates the attachment of Toxoplasma gondii to cell-surface proteoglycans. Mol Biochem Parasitol 2001;116:35-44.
He XL, Grigg ME, Boothroyd JC, Garcia KC. Structure of the immunodominant surface antigen from the Toxoplasma gondii SRS superfamily. Nat Struct Biol 2002;9:606-611.
Vita R, Overton JA, Greenbaum JA, Ponomarenko J, Clark JD, Cantrell JR, et al. The immune epitope database (IEDB) 3.0. Nucleic Acids Res 2015;43(Database issue):D405-12.
Zhang X, Wang J, Liu M, Wang S, Zhang H, Zhao Y. Cloning, expression, purification, and characterization of the catalytic domain of sika deer MMP-13. Protein Expr Purif 2016;127:16-21.
Arezumand R, Mahdian R, Behdani M, Khanahmad H, Langari J, Namvarasl N, et al. Recombinant expression and purification of human placental growth factor 1 and specific camel heavy chain polyclonal antibody preparation. Saudi J Biol Sci 2014;21:35-39.
Zhang NZ, Wang M, Xu Y, Petersen E, Zhu XQ. Recent advances in developing vaccines against Toxoplasma gondii: an update. Expert Rev Vaccines 2015;14:1609-1621.
Cong H, Zhang M, Xin Q, Wang Z, Li Y, Zhao Q, et al. Compound DNA vaccine encoding SAG1/ SAG3 with A2/B subunit of cholera toxin as a genetic adjuvant protects BALB/c mice against Toxoplasma gondii. Parasit Vectors 2013;6:63.
Wang Y, Wang G, Cai J, Yin H. Review on the identification and role of Toxoplasma gondii antigenic epitopes. Parasitol Res 2016;115:459-468.
Hajissa K, Zakaria R, Suppian R, Mohamed Z. Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans. Parasit Vectors 2015;8:315.
| Files | ||
| Issue | Vol 9 No 4 (2017) | |
| Section | Original Article(s) | |
| Keywords | ||
| Toxoplasma gondii Polytopic antigen SAG Epitope mapping | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
